JPH0564928B2 - - Google Patents

Description

[Detailed description of the invention] Stephania tetrandra S.MOORE
is a herbal medicine derived from the roots of the Menispermaceae family, and pharmacological research in China has proven that it has analgesic, anti-inflammatory, antiallergic, and antihypertensive effects. It is known that the roots of Koboshu contain about 1.2% of alkaloids such as tetrandrine, phantinoline, menisin, menisidine, and cyclanoline.

Also, angiotensin
is a decapeptide consisting of 10 amino acids,
It is produced from angiotensinogen, which has a molecular weight of approximately 100,000 and is present in serum, by the enzyme renin, which is present in the kidneys. Furthermore, this angiotensin is converted to active angiotensin by the action of angiotensin converting enzyme. Angiotensin is an octapeptide that has a strong vasoconstrictive effect and is said to be the main renal pressor substance. It is also known that angiotensin converting enzyme degrades and inactivates bradykinin, which has a hypotensive effect. Therefore, angiotensin converting enzyme inhibitors are effective in treating hypertension in that they inhibit the production of pressor substances and the decomposition of antihypertensive substances.

The present inventors conducted research on alkaloids that have angiotensin converting enzyme inhibitory activity among the alkaloid components contained in Kofuji, and found that N,N-dimethyl-phenanthro[3,4-
d]-1,3-dioxole-5-ethylamine has an angiotensin converting enzyme inhibitory effect and has completed the present invention. That is, the present invention is based on the formula An angiotensin converting enzyme inhibitor (hereinafter referred to as the drug of the present invention) containing N,N-dimethyl-phenanthro[3,4-d]-1,3-dioxol-5-ethylamine as an active ingredient. .

For example, the following method can be used to obtain N,N-dimethyl-phenanthro[3,4-d]-1,3-dioxole-5-ethylamine, which is the active ingredient of the drug of the present invention.

Extract Powderfish with water, alcohol, or a mixed solvent of water and alcohol, and remove the solvent from the extract and dissolve the residue as it is or, if necessary, in water or a mixed solvent of water and alcohol. ,
After partitioning with an organic solvent such as n-hexane or petroleum ether to remove fat-soluble components that migrate to the organic solvent, add aqueous ammonia to adjust the pH to 9, and then extract with chloroform and remove the solvent to obtain a chloroform extract. . Next, the chloroform extract was subjected to column chromatography using alumina, developed with chloroform as an eluent, and divided into two fractions. The first fraction was concentrated under reduced pressure and recrystallized using methanol to obtain crystals. It can be obtained by removing tetrandrine, the main alkaloid in the powder, and subjecting the crystal mother liquor to column chromatography using methanol (Sephadex LH20).

N,N-dimethyl-phenanthro[3,4-
A specific example of the production of d]-1,3-dioxole-5-ethylamine is as follows.

Specific example: Extract 7.76Kg of Powdered Powder with 30% methanol, remove methanol from the extract, and make methanol extract.
Obtained 410g. Dissolve this methanol extract in 1.5 ml of a 90% methanol-water mixture and add it to n-hexane.
1.5 to remove fat-soluble components, and then concentrate the 90% methanol-water fraction under reduced pressure. To the resulting residue, add 1.5 of ammonia water with a pH of 9 to dissolve it.
Extraction was performed five times with 1.5 ml of chloroform, and the aqueous ammonia layer and chloroform layer were each concentrated under reduced pressure to obtain an aqueous ammonia extract and a chloroform extract. The chloroform extracted extract was then subjected to alumina (aluminum oxide 90 manufactured by Merck) column chromatography, eluted with chloroform and divided into two fractions, the first fraction was concentrated under reduced pressure and further extracted with tetrandrine using methanol. It crystallized and the crystals were filtered. Next, the crystal mother liquor was subjected to column chromatography (Sephadex LH20) and eluted with methanol.
Rf value 0.38 (thin plate: Kieselgel 60F ₂₅₄ ,
A colorless oily substance containing a developing solvent: chloroform-methanol 3:1 and a coloring reagent: Dragendorff's reagent was obtained. The physical and chemical properties of this substance are described in the literature [ST
Akramov and SYYunusov, Khim.Prir.
Soedin., 4(6) , 390 (1968)] N,N-dimethyl-phenanthro[3,4-d]-1,3-
This was consistent with the physicochemical properties of dioxol-5-ethylamine.

Next, the fact that the compound obtained in the specific example, which is an active ingredient of the drug of the present invention, has an angiotensin converting enzyme inhibitory effect will be explained by giving an experimental example.

Experimental example: Suspend 2g of Rabbit Langacetone powder (manufactured by Sigma) in 30ml of 50mM phosphate buffer (PH8.3), centrifuge at 34000g for 40 minutes, and seal the supernatant in a dialysis tube. After dialyzing with 10mM phosphate buffer 3, the mixture was further appropriately diluted with 50mM phosphate buffer to obtain an angiotensin converting enzyme solution.

The compound obtained in the specific example was dissolved in 20% dimethyl sulfoxide to a final concentration of 1mM, 25μ of this was placed in a test tube, and 50μ of hyperlyl histidyl leucine (final concentration 5mM) was added as a substrate.
and 25μ of 3M sodium chloride solution,
Add 50μ of 500mM phosphate buffer (PH8.3),
After incubating at 37°C for 10 minutes, 100μ of the enzyme solution obtained as described above was added and reacted at 37°C for 60 minutes. After that, 100μ of 1N hydrochloric acid was added to stop the reaction.
Benzoylalanine (1mg/ml) as internal standard
Add 25μ, add 1ml of ethyl acetate, and measure the amount of hipric acid extracted in ethyl acetate by high-performance liquid chromatography [column, μ-Bondapak
C ₁₈ (diameter 4 mm, length 30 cm); mobile phase, acetonitrile: methanol: 1% acetic acid (1:1:8); flow rate,
1 ml/min; detection, ultraviolet light (254 nm)], and this was taken as the enzyme activity.

Regarding this result, the inhibition rate was calculated using the following formula.

Inhibition rate = C-S/C x 100 (%) C: Peak area of hipric acid when the compound obtained in the specific example is not included (corrected by internal standard) S: Hipric acid when the compound obtained in the specific example is added Peak area of acid (corrected by internal standard) This gives an inhibition rate of the compound obtained in the specific example of 61.4
%, and angiotensin converting enzyme inhibitory effect was observed.

Next, an acute toxicity test of the compound obtained in the specific example, which is the active ingredient of the drug of the present invention, was conducted using ddY male mice. There were no examples.

As described above, the compounds obtained in the specific examples have extremely low toxicity and high safety.

In addition, the compounds obtained in the specific examples can also be used as salts commonly used as pharmaceuticals such as hydrochloric acid, sulfuric acid, and succinic acid in order to achieve the desired effects.

Next, the dosage and formulation of the drug of the present invention will be explained.

The compounds obtained in the specific examples can be administered to animals and humans as they are or together with conventional pharmaceutical carriers. The dosage form is not particularly limited and may be selected and used as appropriate, and may include oral preparations such as tablets, capsules, and granules.

In order to exert the desired effect as an oral agent,
Although it varies depending on the patient's age, weight, and severity of the disease,
Normally, one day as the weight of the compound obtained in the specific example for an adult.
It seems appropriate to take 120 to 600 mg in up to three divided doses.

In the present invention, oral preparations such as tablets, capsules, and granules are manufactured according to conventional methods. Tablets are manufactured by mixing and shaping the compound obtained in the specific example with pharmaceutical excipients such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic, etc. Capsules are manufactured by mixing the compound obtained in the specific example with pharmaceutical excipients such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic, etc. It is manufactured by mixing it with an active pharmaceutical filler or diluent and filling it into hard gelatin capsules, soft gelatin capsules, etc. Syrups and elixirs are produced by mixing the compounds obtained in the specific examples with sweeteners such as sucrose, preservatives such as methyl and propylparabens, coloring agents, seasonings, fragrances, and adjuvants.

Next, the present invention will be specifically explained with reference to examples, but the present invention is not limited thereto.

Example 1 Mix 100g of the compound obtained in the specific example with 20g of silicic anhydride, add 75g of corn starch,
Further mixing was performed. To this mixture was added 100 ml of a 10% hydroxypropyl cellulose ethanol solution, and the mixture was stirred in a conventional manner, extruded, dried, and sieved to obtain granules of 20 to 50 mesh particles.

This granule has a dosage of 80 to 400 depending on the symptoms.
mg (40 to 200 mg as the weight of the compound obtained in the specific example)
(equivalent to 3 times a day).

Example 2 Mix 40g of the compound obtained in the specific example with 20g of silicic anhydride, add and mix 10g of microcrystalline cellulose, 0.5g of magnesium stearate, and 49.5g of lactose,
This mixture was compressed using a single-shot tablet machine to produce tablets with a diameter of 7 mm and a weight of 120 mg.

One tablet of the present invention contains 40 mg of the compound obtained in the specific example. This tablet is taken 1 to 5 tablets at a time, three times a day.

Example 3 Mix 40 mg of the compound obtained in the specific example with 100 mg of lactose,
Capsules were obtained by filling No. 0 gelatin capsules.

This capsule preparation is taken 1 to 5 capsules at a time, three times a day, depending on the symptoms.

Claims (1)

[Claims] 1 formula An angiotensin converting enzyme inhibitor containing N,N-dimethyl-phenanthro[3,4-d]-1,3-dioxol-5-ethylamine as an active ingredient.